title: Impact of blood sampling technique on blood quality and animal welfare in haemophilic mice.
abstract: Blood collection in mice can be a challenge, in particular for samples used for coagulation analysis as initiation of coagulation during the procedures can influence the results. Blood collection from the retrobulbar venous plexus is commonly used but the method remains controversial. Several alternatives exist but not all are applicable to mice with a compromised coagulation system because of subsequently excessive bleeding. We therefore wanted to explore whether blood collection by puncture of the submandibular vein could replace blood collected from the retrobulbar venous plexus during pharmacokinetic and pharmacodynamic studies in mice lacking coagulation factor VIII (FVIII). The plasma concentrations of recombinant activated factor VII were independent of the blood collection method in a pharmacokinetic study. The same applied to the thromboelastographic profile of mice with normal coagulation in a pharmacodynamic study. However, excessive haemorrhages were observed in all FVIII knockout mice after a single puncture of the submandibular vein and 60% of the mice were euthanized 2-4 h after the blood collection. In contrast, no or only slight haemorrhage was observed in animals subjected to blood collection from the retrobulbar venous plexus. No signs of distress determined by blood glucose level or clinical abnormalities of the eye were observed after puncture of the retrobulbar venous plexus. In conclusion, blood collected by puncture of the submandibular vein and retrobulbar venous plexus has a quality which allows it to be used in coagulation assays. However, because of excessive bleedings, puncture of the submandibular vein is not recommended in mice lacking FVIII.
authors.affiliation: Haemostasis Pharmacology, Novo Nordisk A/S, Novo Nordisk Park, 2760 Maaloev, Denmark. HLiA@novonordisk.com
authors.forename: Heidi
authors.initials: H
authors.lastname: Holmberg
authors.forename: Maria Kristina
authors.initials: MK
authors.lastname: Kiersgaard
authors.forename: Lars Friis
authors.initials: LF
authors.lastname: Mikkelsen
authors.forename: Mikael
authors.initials: M
authors.lastname: Tranholm
mesh_terms: D000827:Animal Welfare; D000818:Animals; D001777:Blood Coagulation; D001786:Blood Glucose / Q000032:analysis; D001800:Blood Specimen Collection / Q000379:methods / Q000662:veterinary; D005123:Eye / Q000098:blood supply; D005167:Factor VII / Q000008:administration & dosage / Q000493:pharmacokinetics; D005169:Factor VIII / Q000378:metabolism*; D005260:Female; D006470:Hemorrhage; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D013363:Submandibular Gland / Q000098:blood supply; D014680:Veins / Q000293:injuries
chemical_list: D001786:Blood Glucose; D005167:Factor VII; D005169:Factor VIII